1. Observed bromodomain flexibility reveals histone peptide- and small molecule ligand-compatible forms of ATAD2.
- Author
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Poncet-Montange G, Zhan Y, Bardenhagen JP, Petrocchi A, Leo E, Shi X, Lee GR 4th, Leonard PG, Geck Do MK, Cardozo MG, Andersen JN, Palmer WS, Jones P, and Ladbury JE
- Subjects
- ATPases Associated with Diverse Cellular Activities, Adenosine Triphosphatases antagonists & inhibitors, Adenosine Triphosphatases genetics, Adenosine Triphosphatases metabolism, Antineoplastic Agents chemical synthesis, Antineoplastic Agents chemistry, Antineoplastic Agents pharmacology, Binding Sites, Biotinylation, DNA-Binding Proteins antagonists & inhibitors, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Enzyme Inhibitors chemical synthesis, Enzyme Inhibitors pharmacology, Histones antagonists & inhibitors, Histones metabolism, Humans, Isoxazoles chemical synthesis, Isoxazoles pharmacology, Kinetics, Ligands, Mutant Proteins antagonists & inhibitors, Mutant Proteins chemistry, Mutant Proteins metabolism, Peptide Fragments antagonists & inhibitors, Peptide Fragments metabolism, Pliability, Protein Conformation, Protein Interaction Domains and Motifs, Recombinant Proteins chemistry, Recombinant Proteins metabolism, Sulfonamides chemical synthesis, Sulfonamides chemistry, Sulfonamides pharmacology, meta-Aminobenzoates chemical synthesis, meta-Aminobenzoates chemistry, meta-Aminobenzoates pharmacology, Adenosine Triphosphatases chemistry, DNA-Binding Proteins chemistry, Drug Design, Enzyme Inhibitors chemistry, Histones chemistry, Isoxazoles chemistry, Peptide Fragments chemistry, Protein Processing, Post-Translational
- Abstract
Preventing histone recognition by bromodomains emerges as an attractive therapeutic approach in cancer. Overexpression of ATAD2 (ATPase family AAA domain-containing 2 isoform A) in cancer cells is associated with poor prognosis making the bromodomain of ATAD2 a promising epigenetic therapeutic target. In the development of an in vitro assay and identification of small molecule ligands, we conducted structure-guided studies which revealed a conformationally flexible ATAD2 bromodomain. Structural studies on apo-, peptide-and small molecule-ATAD2 complexes (by co-crystallization) revealed that the bromodomain adopts a 'closed', histone-compatible conformation and a more 'open' ligand-compatible conformation of the binding site respectively. An unexpected conformational change of the conserved asparagine residue plays an important role in driving the peptide-binding conformation remodelling. We also identified dimethylisoxazole-containing ligands as ATAD2 binders which aided in the validation of the in vitro screen and in the analysis of these conformational studies.
- Published
- 2015
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